Xuya Yu

Effects of additional Mg2+ on the growth, lipid production, and fatty acid composition of Monoraphidium sp. FXY-10 under different culture conditions

The effects of supplementing the culture medium with Mg2+ on the growth, lipid production, and fatty acid composition of Monoraphidium sp. FXY-10 were studied under photoautotrophic, heterotrophic, and mixotrophic conditions. Under the photoautotrophic condition, microalgae supplemented with 100 μM Mg2+ grew significantly better than the control group and exhibited a secondary growth state. The final cell density was 1.25-fold higher than that of the control group (2.98 g L−1), and the peak lipid content reached 59.8 % (control group 52.3 %). Culture under the heterotrophic condition did not significantly increase the growth rate, but the experimental group (100 μM Mg2+ supplementation) achieved a 37.03 % lipid content compared to 28.47 % by the control group. The lipid productivity of the experimental group (100 μM Mg2+ supplementation) was higher, reaching 65.93 mg L−1 day−1 compared with 56.10 mg L−1 day−1 for the group without additional Mg2+. Under the mixotrophic condition, the experimental group achieved a final density of 3.10 g L−1, which was higher than that of the control group (2.98 g L−1). There was also no variation in fatty acid composition between the experimental group and the control group. Under the heterotrophic and mixotrophic conditions, the experimental group produced more than 50% saturated fatty and mono-unsaturated fatty acids, and the degree of unsaturation was <137. This result was relatively lower than that of the control.

Improved polysaccharide production in a submerged culture of Ganoderma lucidum by the heterologous expression of Vitreoscilla hemoglobin gene.

Expression of Vitreoscilla hemoglobin (VHb) gene was used to improve polysaccharide production in Ganoderma lucidum. The VHb gene, vgb, under the control of the constitutive glyceraldehyde-3-phosphate dehydrogenase gene promoter was introduced into G. lucidum. The activity of expressed VHb was confirmed by the observation of VHb specific CO-difference spectrum with a maximal absorption at 419 nm for the transformant. The effects of VHb expression on intracellular polysaccharide (IPS) content, extracellular polysaccharide (EPS) production and transcription levels of three genes encoding the enzymes involved in polysaccharide biosynthesis, including phosphoglucomutase (PGM), uridine diphosphate glucose pyrophosphorylase (UGP), and β-1,3-glucan synthase (GLS), were investigated. The maximum IPS content and EPS production in the vgb-bearing G. lucidum were 26.4 mg/100mg dry weight and 0.83 g/L, respectively, which were higher by 30.5% and 88.2% than those of the wild-type strain. The transcription levels of PGM, UGP and GLS were up-regulated by 1.51-, 1.55- and 3.83-fold, respectively, in the vgb-bearing G. lucidum. This work highlights the potential of VHb to enhance G. lucidum polysaccharide production by large scale fermentation.

Production of biomass and lipids by the oleaginous microalgae Monoraphidium sp. QLY-1 through heterotrophic cultivation and photo-chemical modulator induction

A two-step strategy comprising heterotrophic cultivation and photo-chemical modulator induction was developed to enhance biomass and lipid accumulation in the oleaginous Monoraphidium sp. QLY-1, which was isolated from Qilu Lake in Yunnan Plateau. The algae were first cultivated heterotrophically to achieve high biomass concentration (5.54gL(-1)) with a lipid content of 22.47%. The cultivated algae were diluted, transferred to light environment, and treated with different chemical elicitors. Results showed that the lipid content increased to 36.68% after 3-day of photoinduction. The lipid content was further enhanced by 1.21, 1.32, and 1.29 folds in algal cells treated with nitrogen deficiency, 20gL(-1) NaCl, and 5mM glycine betaine, respectively. The maximum lipid content (48.54%) and lipid productivity (121.27mgL(-1)d(-1)) were obtained in treatments with 20gL(-1) NaCl and 5mM GB, respectively. This study proposes a strategy to efficiently produce lipids by using microalgae.

Effect of fulvic acid induction on the physiology, metabolism, and lipid biosynthesis-related gene transcription of Monoraphidium sp. FXY-10

Fulvic acid (FA) triggers lipid accumulation in Monoraphidium sp. FXY-10, which can produce biofuels. Therefore, the metabolism shift and gene expression changes influenced by fulvic acid should be investigated. In this study, lipid and protein contents increased rapidly from 44.6% to 54.3% and from 31.4% to 39.7% under FA treatment, respectively. By contrast, carbohydrate content sharply declined from 49.5% to 32.5%. The correlation between lipid content and gene expression was also analyzed. Results revealed that accD, ME, and GPAT genes were significantly correlated with lipid accumulation. These genes could likely influence lipid accumulation and could be selected as modification candidates. These results demonstrated that FA significantly increased microalgal lipid accumulation by changing the intracellular reactive oxygen species, gene expression, and enzyme activities of acetyl-CoA carboxylase, malic enzyme, and phosphoenolpyruvate carboxylase.

A strategy for promoting lipid production in green microalgae Monoraphidium sp. QLY-1 by combined melatonin and photoinduction

Microalgae lipids are potential candidates for the production of renewable biodiesel. The combination of plant hormones and two-step cultivation regulates lipid production in microalgae. A strategy for promoting lipid accumulation in Monoraphidium sp. QLY-1 by combining exogenous melatonin (MT) and photoinduction was developed. The effects of melatonin on the lipid content, reactive oxygen species (ROS), and activities of three key fatty acid biosynthetic enzyme in Monoraphidium sp. QLY-1 were investigated. The lipid content increased by 1.32-fold under 1μM melatonin treatment. The maximum lipid content achieved was 49.6%. However, the protein and carbohydrate contents decreased rapidly from 57.21% to 47.96% and from 53.4% to 37.71%, respectively. Biochemical and physiological analyses suggested that the ROS and lipid biosynthesis-related enzyme activities correlated with increased lipid accumulation under photo-melatonin induction conditions.

Enhanced astaxanthin production from Haematococcus pluvialis using butylated hydroxyanisole

Haematococcus pluvialis is a promising natural source of high-value antioxidant astaxanthin under stress conditions. Biotic or abiotic elicitors are effective strategies for improving astaxanthin production in H. pluvialis. Butylated hydroxyanisole (BHA) was identified as an effective inducer for H. pluvialis LUGU. Under a treatment of 2mgL(-1) BHA (BHA2), astaxanthin content reached a maximum of 29.03mgg(-1) dry weight (DW) (2.03-fold of that in the control) after 12day of the mid-exponential growth phase. Subsequently, H. pluvialis LUGU was subjected to BHA2 at different growth phases because an appropriate time node for adding elicitors is vital for the entire production to succeed. As a result, the highest astaxanthin content (29.3mgg(-1) DW) was obtained in cells on day 14 (BHA2 14) of the late-exponential growth phase. Furthermore, the samples treated with BHA2 14 and the control group were compared in terms of the transcriptional expression of seven carotenogenesis genes, fatty acid composition, and total accumulated astaxanthin. All selected genes exhibited up-regulated expression profiles, with chy, crtO, and bkt exhibiting higher maximum transcriptional levels than the rest. Oleic acid content increased 33.15-fold, with acp, fad, and kas expression being enhanced on the day when astaxanthin was produced rapidly.

Melatonin enhances lipid production in Monoraphidium sp. QLY-1 under nitrogen deficiency conditions via a multi-level mechanism.

In this study, melatonin (MT) promoted lipid accumulation in Monoraphidium sp. QLY-1 under nitrogen deficiency conditions. The lipid accumulation increased 1.22- and 1.36-fold compared with a nitrogen-starved medium and a normal BG-11 medium, respectively. The maximum lipid content was 51.38%. The reactive oxygen species (ROS) level in the presence of melatonin was lower than that in the control group, likely because of the high antioxidant activities. The application of melatonin upregulated the gibberellin acid (GA) production and rbcL and accD expression levels but downregulated the abscisic acid (ABA) content and pepc expression levels. These findings demonstrated that exogenous melatonin could further improve the lipid production in Monoraphidium sp. QLY-1 by regulating antioxidant systems, signalling molecules, and lipid biosynthesis-related gene expression under nitrogen deficiency conditions.

Overexpression of the Squalene Epoxidase Gene Alone and in Combination with the 3-Hydroxy-3-methylglutaryl Coenzyme A Gene Increases Ganoderic Acid Production in Ganoderma lingzhi

The squalene epoxidase (SE) gene from the biosynthetic pathway of ganoderic acid (GA) was cloned and overexpressed in Ganoderma lingzhi. The strain that overexpressed the SE produced approximately 2 times more GA molecules than the wild-type (WT) strain. Moreover, SE overexpression upregulated lanosterol synthase gene expression in the biosynthetic pathway. These results indicated that SE stimulates GA accumulation. Then, the SE and 3-hydroxy-3-methylglutaryl coenzyme A (HMGR) genes were simultaneously overexpressed in G. lingzhi. Compared with the individual overexpression of SE or HMGR, the combined overexpression of the two genes further enhanced individual GA production. The overexpressing strain produced maximum GA-T, GA-S, GA-Mk, and GA-Me contents of 90.4 ± 7.5, 35.9 ± 5.4, 6.2 ± 0.5, and 61.8 ± 5.8 μg/100 mg dry weight, respectively. These values were 5.9, 4.5, 2.4, and 5.8 times higher than those produced by the WT strain. This is the first example of the successful manipulation of multiple biosynthetic genes to improve GA content in G. lingzhi.

Melatonin: A Multifunctional Molecule That Triggers Defense Responses against High Light and Nitrogen Starvation Stress in Haematococcus pluvialis

Melatonin (MLT), a ubiquitously distributed small molecule, functions in plant responses to various biotic and abiotic stresses. However, the interactions between melatonin and other important molecules in Haematococcus pluvialis response stresses are largely unknown. In the present study, exogenous melatonin improved H. pluvialis resistance to nitrogen starvation and high light. We concluded that exogenous melatonin treatment prevented the reactive oxygen species (ROS) burst and limited cell damage induced by abiotic stress through activation of antioxidant enzymes and antioxidants. Astaxanthin, a major antioxidant in H. pluvialis cells, exhibited a 2.25-fold increase in content after treatment with melatonin. The maximal astaxanthin content was 32.4 mg g-1. The functional roles of the nitric oxide (NO)-mediated mitogen activated protein kinase (MAPK) signaling pathway and cyclic adenosine monophosphate (cAMP) signaling pathway induced by melatonin were also evaluated. The results clearly indicate that cAMP signaling pathways are positively associated with microalgal astaxanthin biosynthesis. Additionally, the NO-dependent MAPK signaling cascade is activated in response to astaxanthin accumulation induced by melatonin, confirming that MAPK is a target of NO action in physiological processes. This work is the first to use H. pluvialis as in vivo model and documents the influence of melatonin on the physiological response to abiotic stress in this microalgae.

Butylated hydroxytoluene induces astaxanthin and lipid production in Haematococcus pluvialis under high-light and nitrogen-deficiency conditions

The aim of this study was to investigate the effects of butylated hydroxytoluene (BHT) on the production of astaxanthin and lipids in Haematococcus pluvialis LUGU under high-light and nitrogen-deficiency conditions. Astaxanthin and lipid contents were increased by 71.13% and 10.71%, respectively, in algal cells treated with 2 mg L-1 BHT. The maximal contents of astaxanthin and lipids were 3.17% and 46%, respectively. The levels of reactive oxygen species (ROS) in the presence of BHT were lower than in the control, and this effect involved strong activation of several antioxidases. Additionally, BHT application upregulated endogenous nitric oxide (NO) production. These results showed that this approach is useful for stimulating production of astaxanthin and lipids in H. pluvialis and that exogenous BHT induces astaxanthin and lipid production, which is responsible for the signalling molecule responses against abiotic stress conditions in H. pluvialis.